VANILREP1 polynucleotides and VANILREP1 polypeptides

ABSTRACT

VANILREP1 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing VANILREP1 polypeptides and polynucleotides in therapy, and diagnostic assays for such.

FIELD OF THE INVENTION

This invention relates to newly identified polypeptides andpolynucleotides encoding such polypeptides, to their use in therapy andin identifying compounds which may be agonists, antagonists and/orinhibitors which are potentially useful in therapy, and to production ofsuch polypeptides and polynucleotides.

BACKGROUND OF THE INVENTION

The drug discovery process is currently undergoing a fundamentalrevolution as it embraces ‘functional genomics’, that is, highthroughput genome- or gene-based biology. This approach as a means toidentify genes and gne products as therapeutic targets is rapidlysuperceding earlier approaches based on ‘positional cloning’. Aphenotype, that is a biological function or genetic disease, would beidentified and this would then be tracked back to the responsible gene,based on its genetic map position.

Functional genomics relies heavily on high-throughput DNA sequencingtechnologies and the various tools of bioinformatics to identify genesequences of potential interest from the many molecular biologydatabases now available. There is a continuing need to identify andcharacterise further genes and their related polypeptides/proteins, astargets for drug discovery.

SUMMARY OF THE INVENTION

The present invention relates to VANILREP1, in particular VANILREP1polypeptides and VANILREP1 polynucleotides, recombinant materials andmethods for their production. In another aspect, the invention relatesto methods for using such polypeptides and polynucleotides, includingthe treatment of pain, chronic pain, neuropathic pain, postoperativepain, rheumatoid arhritic pain, neuralgia, neuropathies, algesia, nerveinjury, ischaemia, neurodegeneration, stroke, incontinence andinflammatory disorders, hereinafter referred to as “the Diseases”,amongst others. In a further aspect, the invention relates to methodsfor identifying agonists and antagonists/inhibitors using the materialsprovided by the invention, and treating conditions associated withVANILREP1 imbalance with the identified compounds. In a still furtheraspect, the invention relates to diagnostic assays for detectingdiseases associated with inappropriate VANILREP1 activity or levels.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows currents evoked from a cell in response to capsaicin over arange of concentrations.

FIG. 2 shows inhibition of the capsaicin evoked response (1 uM) bycapsazepine (10 uM). The inhibition is reversible with extended washing.

DESCRIPTION OF THE INVENTION

In a first aspect, the present invention relates to VANILREP1polypeptides. These include the polypeptide of SEQ ID NO:2 andpolymorphic variants thereof, for example PVP-1, the polypeptide of SEQID NO:8. Such peptides include isolated polypeptides comprising an aminoacid sequence which has at least 90% identity, preferably at least 95%identity, more preferably at least 97-99% identity, to that of SEQ IDNO:2 or SEQ ID NO:8 over the entire length of SEQ ID NO:2 or SEQ ID NO:8respectively. Such polypeptides include those comprising the amino acidof SEQ ID NO:2 or SEQ ID NO:8.

Further peptides of the present invention include isolated polypeptidesin which the amino acid sequence has at least 90% identity, preferablyat least 95% identity, more preferably at least 97-99% identity, to theamino acid sequence of SEQ ID NO:2 or SEQ ID NO:8, over the entirelength of SEQ ID NO:2 or SEQ ID NO:8. Such polypeptides include thepolypeptide of SEQ ID NO:2 and SEQ ID NO:8.

Further peptides of the present invention include isolated polypeptidesencoded by a polynucleotide comprising the sequence contained in SEQ IDNO:1 or SEQ ID NO:7.

Polypeptides of the present invention are believed to be members of theion channel family of polypeptides. They are therefore of interestbecause they are associated with the mechanism of action of capsaicin (avanilloid compound), a constituent of chilli peppers. Capsaicin elicitsa senstation of burning pain by selectively activating sensory neuronsthat convey information about noxious stimuli to the central nervoussystem. The channels are permeable to cations and exhibit a notablepreferance for divalent cations, particularly calcium ions. The level ofcalcium ion permeability exceeds that observed for most non-selectivecation channels and is similar to values observed for NMDA-typeglutamate receptors and alpha7 nicotinic acetylcholine receptors, bothof which are noted for this property. These properties are hereinafterreferred to as VANILREP1 activity” or VANILREP1 polypeptide activity” or“biological activity of VANILREP1”. Also included amongst theseactivities are antigenic and immunogenic activities of said VANILREP1polypeptides, in particular the antigenic and immunogenic activities ofthe polypeptides of SEQ ID NO:2 or SEQ ID NO:8. Preferably, apolypeptide of the present invention exhibits at least one biologicalactivity of VANILREP1.

The polypeptides of the present invention may be in the form of the“mature” protein or may be a part of a larger protein such as aprecursor or a fusion protein. It is often advantageous to include anadditional amino acid sequence which contains secretory or leadersequences, pro-sequences, sequences which aid in purification such asmultiple histidine residues, or an additional sequence for stabilityduring recombinant production.

The present invention also includes variants of the aforementionedpolypeptides, that is polypeptides that vary from the referents byconservative amino acid substitutions, whereby a residue is substitutedby another with like characteristics. Typical such substitutions areamong Ala, Val, Leu and Ile; among Ser and Thr; among the acidicresidues Asp and Glu; among Asn and Gln; and among the basic residuesLys and Arg; or aromatic residues Phe and Tyr. Particularly preferredare variants in which several, 5-10, 1-5, 1-3, 1-2 or 1 amino acids aresubstituted, deleted, or added in any combination.

Polypeptides of the present invention can be prepared in any suitablemanner. Such polypeptides include isolated naturally occurringpolypeptides, recombinantly produced polypeptides, syntheticallyproduced polypeptides, or polypeptides produced by a combination ofthese methods. Means for preparing such polypeptides are well understoodin the art.

In a further aspect, the present invention relates to VANILREP1polynucleotides. Such polynucleotides include isolated polynucleotidescomprising a nucleotide sequence encoding a polypeptide which has atleast 90% identity, preferably at least 95% identity, to the amino acidsequence of SEQ ID NO:2 or SEQ ID NO:8, over the entire length of SEQ IDNO:2 or SEQ ID NO:8, respectively. In this regard, polypeptides whichhave at least 97% identity are highly preferred, whilst those with atleast 98-99% identity are more highly preferred, and those with at least99% identity are most highly preferred. Such polynucleotides include apolynucleotide comprising the nucleotide sequence contained in SEQ IDNO:1 or SEQ ID NO:7, encoding the polypeptide of SEQ ID NO:2 or SEQ IDNO:8, respectively.

Further polynucleotides of the present invention include isolatedpolynucleotides comprising a nucleotide sequence that has at least 85%identity, preferably at least 90% identity, more preferably at least 95%identity, to a nucleotide sequence encoding a polypeptide of SEQ ID NO:2or SEQ ID NO:8, respectively, over the entire coding region. In thisregard, polynucleotides which have at least 97% identity are highlypreferred, whilst those with at least 98-99% identity are more highlypreferred, and those with at least 99% identity are most highlypreferred.

Further polynucleotides of the present invention include isolatedpolynucleotides comprising a nucleotide sequence which has at least 85%identity, preferably at least 90% identity, more preferably at least 95%identity, to SEQ ID NO:1 or SEQ ID NO:7, over the entire length of SEQID NO:1 or SEQ ID NO:7, respectively. In this regard, polynucleotideswhich have at least 97% identity are highly preferred, whilst those withat least 98-99% identity are more highly preferred, and those with atleast 99% identity are most highly preferred. Such polynucleotidesinclude a polynucleotide comprising the polynucleotide of SEQ ID NO:1 orSEQ ID NO:7, as well as the polynucleotide of SEQ ID NO:1 and SEQ IDNO:7, respectively.

The invention also provides polynucleotides which are complementary toall the above described polynucleotides.

The nucleotide sequences of SEQ ID NO:1 and SEQ ID NO:7 show homologywith rat vanilloid receptor VR1(M. J. Caterina et al., Nature 389:816-824, 1997). The nucleotide sequence of SEQ ID NO:1 is a cDNAsequence and comprises a polypeptide encoding sequence (nucleotide 864to 3380) encoding a polypeptide of 839 amino acids, the polypeptide ofSEQ ID NO:2. The nucleotide sequence encoding the polypeptide of SEQ IDNO:2 may be identical to the polypeptide encoding sequence contained inSEQ ID NO:1 or it may be a sequence other than the one contained in SEQID NO:1, which, as a result of the redundancy (degeneracy) of thegenetic code, also encodes the polypeptide of SEQ ID NO:2. Thepolypeptide of the SEQ ID NO:2 is structurally related to other proteinsof the ion channel family, having homology and/or structural similaritywith rat vanilloid receptor VR1 (M. J. Caterina et al., Nature 389:816-824, 1997).

The nucleotide sequence of SEQ ID NO:7 is a cDNA sequence and comprisesa polypeptide encoding sequence (nucleotide 864 to 3380) encoding apolypeptide of 839 amino acids, the polypeptide of SEQ ID NO:8. Thenucleotide sequence encoding the polypeptide of SEQ ID NO:8 may beidentical to the polypeptide encoding sequence contained in SEQ ID NO:7or it may be a sequence other than the one contained in SEQ ID NO:7,which, as a result of the redundancy (degeneracy) of the genetic code,also encodes the polypeptide of SEQ ID NO:8. The polypeptide of the SEQID NO:8 is structurally related to other proteins of the ion channelfamily, having homology and/or structural similarity with rat vanilloidreceptor VR1(M. J. Caterina et al., Nature 389: 816-824, 1997).

Preferred polypeptides and polynucleotides of the present invention areexpected to have, inter alia, similar biological functions/properties totheir homologous polypeptides and polynucleotides. Furthermore,preferred polypeptides and polynucleotides of the present invention haveat least one VANILREP1 activity.

The present invention also relates to partial or other polynucleotideand polypeptide sequences which were first identified prior to thedetermination of the corresponding full length sequences of SEQ ID NO:1,SEQ ID NO:7, SEQ ID NO:2 and SEQ ID NO:8.

Accordingly, in a further aspect, the present invention provides for anisolated polynucleotide which:

(a) comprises a nucleotide sequence which has at least 85% identity,preferably at least 90% identity, more preferably at least 95% identity,yet more preferably at least 97-99% identity to SEQ ID NO:3 or SEQ IDNO:5, over the entire length of SEQ ID NO:3 or SEQ ID NO:5,respectively;

(b) has a nucleotide sequence which has at least 85% identity,preferably at least 90% identity, more preferably at least 95% identity,yet more preferably at least 97-99% identity, to SEQ ID NO:3 or SEQ IDNO:5, over the entire length of SEQ ID NO:3 or SEQ ID NO:5,respectively;

(c) the polynucleotide of SEQ ID NO:3 or SEQ ID NO:5; or

(d) a nucleotide sequence encoding a polypeptide which has at least 90%identity, preferably at least 95% identity, more preferably at least97-99% identity, to the amino acid sequence of SEQ ID NO:4 or SEQ IDNO:6, over the entire length of SEQ ID NO:4 or SEQ ID NO:6,respectively;

as well as the polynucleotides of SEQ ID NO:3 and SEQ ID NO:5.

The present invention further provides for a polypeptide which:

(a) comprises an amino acid sequence which has at least 90% identity,preferably at least 95% identity, more preferably at least 97-99%identity, to that of SEQ ID NO:4 or SEQ ID NO:6, over the entire lengthof SEQ ID NO:4 or SEQ ID NO:6, respectively;

(b) has an amino acid sequence which has at least 90% identity,preferably at least 95% identity, more preferably at least 97-99%identity, to the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:6, overthe entire length of SEQ ID NO:4 or SEQ ID NO:6, respectively;

(c) comprises the amino acid of SEQ ID NO:4 or SEQ ID NO:6; and

(d) is the polypeptide of SEQ ID NO:4 or SEQ ID NO:6;

as well as polypeptides encoded by a polynucleotide comprising thesequence contained in SEQ ID NO:3 or SEQ ID NO:5.

The nucleotide sequences of SEQ ID NO:3 and SEQ ID NO:5, and the peptidesequences encoded thereby are derived from EST (Expressed Sequence Tag)sequences. It is recognised by those skilled in the art that there willinevitably be some nucleotide sequence reading errors in EST sequences(see Adams, M. D. et al, Nature 377 (supp) 3, 1995). Accordingly, thenucleotide sequences of SEQ ID NO:3 and SEQ ID NO:5, and the peptidesequence encoded therefrom, are therefore subject to the same inherentlimitations in sequence accuracy.

Polynucleotides of the present invention may be obtained, using standardcloning and screening techniques, from a cDNA library derived from mRNAin cells of human brain, cerebellum, dorsal root ganglia, thymus,leukocytes, placenta, foetal liver spleen and ovary, using the expressedsequence tag (EST) analysis (Adams, M. D., et al. Science (1991)252:1651-1656; Adams, M. D. et al., Nature, (1992) 355:632-634; Adams,M. D., et al., Nature (1995) 377 Supp:3-174). Polynucleotides of theinvention can also be obtained from natural sources such as genomic DNAlibraries or can be synthesized using well known and commerciallyavailable techniques.

When polynucleotides of the present invention are used for therecombinant production of polypeptides of the present invention, thepolynucleotide may include the coding sequence for the maturepolypeptide, by itself; or the coding sequence for the maturepolypeptide in reading frame with other coding sequences, such as thoseencoding a leader or secretory sequence, a pre-, or pro- orprepro-protein sequence, or other fusion peptide portions. For example,a marker sequence which facilitates purification of the fusedpolypeptide can be encoded. In certain preferred embodiments of thisaspect of the invention, the marker sequence is a hexa-histidinepeptide, as provided in the pQE vector (Qiagen, Inc.) and described inGentz et al., Proc Natl Acad Sci USA (1989) 86:821-824, or is an HA tag.The polynucleotide may also contain noncoding 5′ and 3′ sequences, suchas transcribed, non-translated sequences, splicing and polyadenylationsignals, ribosome binding sites and sequences that stabilize mRNA.

Further embodiments of the present invention include polynucleotidesencoding polypeptide variants which comprise the amino acid sequence ofSEQ ID NO:2 or SEQ ID NO:8, respectively and in which several, forinstance from 5 to 10, 1 to 5, 1 to 3, 1 to 2 or 1, amino acid residuesare substituted, deleted or added, in any combination.

Polynucleotides which are identical or sufficiently identical to anucleotide sequence contained in SEQ ID NO:1 or SEQ ID NO:7,respectively, may be used as hybridization probes for cDNA and genomicDNA or as primers for a nucleic acid amplification (PCR) reaction, toisolate full-length cDNAs and genomic clones encoding polypeptides ofthe present invention and to isolate cDNA and genomic clones of othergenes (including genes encoding paralogs from human sources andorthologs and paralogs from species other than human) that have a highsequence similarity to SEQ ID NO:1 or SEQ ID NO:7,. Typically thesenucleotide sequences are 70% identical, preferably 80% identical, morepreferably 90% identical, most preferably 95% identical to that of thereferent. The probes or primers will generally comprise at least 15nucleotides, preferably, at least 30 nucleotides and may have at least50 nucleotides. Particularly preferred probes will have between 30 and50 nucleotides. Particularly preferred primers will have between 20 and25 nucleotides.

A polynucleotide encoding a polypeptide of the present invention,including homologs from species other than human, may be obtained by aprocess which comprises the steps of screening an appropriate libraryunder stringent hybridization conditions with a labeled probe having thesequence of SEQ ID NO:1 or SEQ ID NO:7, respectively or a fragmentthereof; and isolating full-length cDNA and genomic clones containingsaid polynucleotide sequence. Such hybridization techniques are wellknown to the skilled artisan. Preferred stringent hybridizationconditions include overnight incubation at 42° C. in a solutioncomprising: 50% formamide, 5× SSC (150 mM NaCl, 15 mM trisodiumcitrate), 50 mM sodium phosphate (pH7.6), 5× Denhardt's solution, 10%dextran sulfate, and 20 microgram/ml denatured, sheared salmon spermDNA; followed by washing the filters in 0.1× SSC at about 65° C. Thusthe present invention also includes polynucleotides obtainable byscreening an appropriate library under stingent hybridization conditionswith a labeled probe having the sequence of SEQ ID NO:1 or SEQ ID NO:7,or a fragment thereof.

The skilled artisan will appreciate that, in many cases, an isolatedcDNA sequence will be incomplete, in that the region coding for thepolypeptide is short at the 5′ end of the cDNA. This is a consequence ofreverse transcriptase, an enzyme with inherently low ‘processivity’ (ameasure of the ability of the enzyme to remain attached to the templateduring the polymerisation reaction), failing to complete a DNA copy ofthe mRNA template during 1st strand cDNA synthesis.

There are several methods available and well known to those skilled inthe art to obtain full-length cDNAs, or extend short cDNAs, for examplethose based on the method of Rapid Amplification of cDNA ends (RACE)(see, for example, Frohman et al., PNAS USA 85, 8998-9002, 1988). Recentmodifications of the technique, exemplified by the Marathon™ technology(Clontech Laboratories Inc.) for example, have significantly simplifiedthe search for longer cDNAs. In the Marathon™ technology, cDNAs havebeen prepared from mRNA extracted from a chosen tissue and an ‘adaptor’sequence ligated onto each end. Nucleic acid amplification (PCR) is thencarried out to amplify the ‘missing’ 5′ end of the cDNA using acombination of gene specific and adaptor specific oligonucleotideprimers. The PCR reaction is then repeated using ‘nested’ primers, thatis, primers designed to anneal within the amplified product (typicallyan adaptor specific primer that anneals further 3′ in the adaptorsequence and a gene specific primer that anneals further 5′ in the knowngene sequence). The products of this reaction can then be analysed byDNA sequencing and a full-length cDNA constructed either by joining theproduct directly to the existing cDNA to give a complete sequence, orcarrying out a separate full-length PCR using the new sequenceinformation for the design of the 5′ primer.

Recombinant polypeptides of the present invention may be prepared byprocesses well known in the art from genetically engineered host cellscomprising expression systems. Accordingly, in a further aspect, thepresent invention relates to expression systems which comprise apolynucleotide or polynucleotides of the present invention, to hostcells which are genetically engineered with such expression sytems andto the production of polypeptides of the invention by recombinanttechniques. Cell-free translation systems can also be employed toproduce such proteins using RNAs derived from the DNA constructs of thepresent invention.

For recombinant production host cells can be genetically engineered toincorporate expression systems or portions thereof for polynucleotidesof the present invention. Introduction of polynucleotides into hostcells can be effected by methods described in many standard laboratorymanuals, such as Davis et al, Basic Methods in Molecular Biology (1986)and Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed.,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989).Preferred such methods include, for instance, calcium phosphatetransfection, DEAE-dextran mediated transfection, transvection,microinjection, cationic lipid-mediated transfection, electroporation,transduction, scrape loading, ballistic introduction or infection.

Representative examples of appropriate hosts include bacterial cells,such as Streptococci, Staphylococci, E. coli, Streptomyces and Bacillussubtilis cells; fungal cells, such as yeast cells and Aspergillus cells;insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animalcells such as CHO, COS, HeLa, C127, 3T3, BHK, HEK 293 and Bowes melanomacells; and plant cells.

A great variety of expression systems can be used, for instance,chromosomal, episomal and virus-derived systems, e.g., vectors derivedfrom bacterial plasmids, from bacteriophage, from transposons, fromyeast episomes, from insertion elements, from yeast chromosomalelements, from viruses such as baculoviruses, papova viruses, such asSV40, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabiesviruses and retroviruses, and vectors derived from combinations thereof,such as those derived from plasmid and bacteriophage genetic elements,such as cosmids and phagemids. The expression systems may containcontrol regions that regulate as well as engender expression. Generally,any system or vector which is able to maintain, propagate or express apolynucleotide to produce a polypeptide in a host may be used. Theappropriate nucleotide sequence may be inserted into an expressionsystem by any of a variety of well-known and routine techniques, suchas, for example, those set forth in Sambrook et al., Molecular Cloning,A Laboratory Manual (supra). Appropriate secretion signals may beincorporated into the desired polypeptide to allow secretion of thetranslated protein into the lumen of the endoplasmic reticulum, theperiplasmic space or the extracellular environment. These signals may beendogenous to the polypeptide or they may be heterologous signals.

If a polypeptide of the present invention is to be expressed for use inscreening assays, it is generally preferred that the polypeptide beproduced at the surface of the cell. In this event, the cells may beharvested prior to use in the screening assay. If the polypeptide issecreted into the medium, the medium can be recovered in order torecover and purify the polypeptide. If produced intracellularly, thecells must first be lysed before the polypeptide is recovered.

Polypeptides of the present invention can be recovered and purified fromrecombinant cell cultures by well-known methods including ammoniumsulfate or ethanol precipitation, acid extraction, anion or cationexchange chromatography, phosphocellulose chromatography, hydrophobicinteraction chromatography, affinity chromatography, hydroxylapatitechromatography and lectin chromatography. Most preferably, highperformance liquid chromatography is employed for purification. Wellknown techniques for refolding proteins may be employed to regenerateactive conformation when the polypeptide is denatured duringintracellular synthesis, isolation and or purification.

This invention also relates to the use of polynucleotides of the presentinvention as diagnostic reagents. Detection of a mutated form of thegene characterised by the polynucleotide of SEQ ID NO:1 or SEQ ID NO:7,respectively which is associated with a dysfunction will provide adiagnostic tool that can add to, or define, a diagnosis of a disease, orsusceptibility to a disease, which results from under-expression,over-expression or altered spatial or temporal expression of the gene.Individuals carrying mutations in the gene may be detected at the DNAlevel by a variety of techniques.

Nucleic acids for diagnosis may be obtained from a subject's cells, suchas from blood, urine, saliva, tissue biopsy or autopsy material. Thegenomic DNA may be used directly for detection or may be amplifiedenzymatically by using PCR or other amplification techniques prior toanalysis. RNA or cDNA may also be used in similar fashion. Deletions andinsertions can be detected by a change in size of the amplified productin comparison to the normal genotype. Point mutations can be identifiedby hybridizing amplified DNA to labeled VANILREP1 nucleotide sequences.Perfectly matched sequences can be distinguished from mismatchedduplexes by RNase digestion or by differences in melting temperatures.DNA sequence differences may also be detected by alterations inelectrophoretic mobility of DNA fragments in gels, with or withoutdenaturing agents, or by direct DNA sequencing (ee, e.g., Myers et al.,Science (1985) 230:1242). Sequence changes at specific locations mayalso be revealed by nuclease protection assays, such as RNase and S1protection or the chemical cleavage method (see Cotton et al., Proc NatlAcad Sci USA (1985) 85:4397-4401). In another embodiment, an array ofoligonucleotides probes comprising VANILREP1 nucleotide sequence orfragments thereof can be constructed to conduct efficient screening ofe.g., genetic mutations. Array technology methods are well known andhave general applicability and can be used to address a variety ofquestions in molecular genetics including gene expression, geneticlinkage, and genetic variability (see for example: M. Chee et al.,Science, Vol 274, pp 610-613 (1996)).

The diagnostic assays offer a process for diagnosing or determining asusceptibility to the Diseases through detection of mutation in theVANILREP1 gene by the methods described. In addition, such diseases maybe diagnosed by methods comprising determining from a sample derivedfrom a subject an abnormally decreased or increased level of polypeptideor MRNA. Decreased or increased expression can be measured at the RNAlevel using any of the methods well known in the art for thequantitation of polynucleotides, such as, for example, nucleic acidamplification, for instance PCR, RT-PCR, RNase protection, Northernblotting and other hybridization methods. Assay techniques that can beused to determine levels of a protein, such as a polypeptide of thepresent invention, in a sample derived from a host are well-known tothose of skill in the art. Such assay methods include radioimmunoassays,competitive-binding assays, Western Blot analysis and ELISA assays.

Thus in another aspect, the present invention relates to a diagonostickit which comprises:

(a) a polynucleotide of the present invention, preferably the nucleotidesequence of SEQ ID NO:1 or SEQ ID NO:7, respectively or a fragmentthereof;

(b) a nucleotide sequence complementary to that of (a);

(c) a polypeptide of the present invention, preferably the polypeptideof SEQ ID NO:2 or SEQ ID NO:8, respectively or a fragment thereof; or

(d) an antibody to a polypeptide of the present invention, preferably tothe polypeptide of SEQ ID NO:2 or SEQ ID NO:8, respectively.

It will be appreciated that in any such kit, (a), (b), (c) or (d) maycomprise a substantial component. Such a kit will be of use indiagnosing a disease or suspectability to a disease, particularly pain,chronic pain, neuropathic pain, postoperative pain, rheumatoid arthriticpain, neuralgia, neuropathies, algesia, nerve injury, ischaemia,neurodegeneration, stroke, incontinence and inflammatory disorders,amongst others.

The nucleotide sequences of the present invention are also valuable forchromosome localisation. The sequence is specifically targeted to, andcan hybridize with, a particular location on an individual humanchromosome. The mapping of relevant sequences to chromosomes accordingto the present invention is an important first step in correlating thosesequences with gene associated disease. Once a sequence has been mappedto a precise chromosomal location, the physical position of the sequenceon the chromosome can be correlated with genetic map data. Such data arefound in, for example, V. McKusick, Mendelian Inheritance in Man(available on-line through Jolms Hopkins University Welch MedicalLibrary). The relationship between genes and diseases that have beenmapped to the same chromosomal region are then identified throughlinkage analysis (coinheritance of physically adjacent genes).

The differences in the cDNA or genomic sequence between affected andunaffected individuals can also be determined. If a mutation is observedin some or all of the affected individuals but not in any normalindividuals, then the mutation is likely to be the causative agent ofthe disease.

The gene of the present invention maps to human chromosome 17p13. Thenucleotide sequences of the present invention are also valuable fortissue localisation. Such techniques allow the determination ofexpression patterns of the human VANILREP1 polypeptides in tissues bydetection of the mRNAs that encode them. These techniques include insitu hybridziation techniques and nucleotide amplification techniques,for example PCR. Such techniques are well known in the art. Results fromthese studies provide an indication of the normal functions of thepolypeptides in the organism. In addition, comparative studies of thenormal expression pattern of human VANILREP1 mRNAs with that of mRNAsencoded by a human VANILREP1 gene provide valuable insights into therole of mutant human VANILREP1 polypeptides, or that of inappropriateexpression of normal human VANILREP1 polypeptides, in disease. Suchinappropriate expression may be of a temporal, spatial or simplyquantitative nature.

The polypeptides of the invention or their fragments or analogs thereof,or cells expressing them, can also be used as immunogens to produceantibodies immunospecific for polypeptides of the present invention. Theterm “immunospecific” means that the antibodies have substantiallygreater affinity for the polypeptides of the invention than theiraffinity for other related polypeptides in the prior art.

Antibodies generated against polypeptides of the present invention maybe obtained by administering the polypeptides or epitope-bearingfragments, analogs or cells to an animal, preferably a non-human animal,using routine protocols. For preparation of monoclonal antibodies, anytechnique which provides antibodies produced by continuous cell linecultures can be used. Examples include the hybridoma technique (Kohler,G. and Milstein, C., Nature (1975) 256:495-497), the trioma technique,the human B-ell hybridoma technique (Kozbor et al., Immunology Today(1983) 4:72) and the EBV-hybridoma technique (Cole et al., MonoclonalAntibodies and Cancer Therapy, 77-96, Alan R. Liss, Inc., 1985).

Techniques for the production of single chain antibodies, such as thosedescribed in U.S. Pat. No. 4,946,778, can also be adapted to producesingle chain antibodies to polypeptides of this invention. Also,transgenic mice, or other organisms, including other mammals, may beused to express humanized antibodies.

The above-described antibodies may be employed to isolate or to identifyclones expressing the polypeptide or to purify the polypeptides byaffinity chromatography.

Antibodies against polypeptides of the present invention may also beemployed to treat the Diseases, amongst others.

In a further aspect, the present invention relates to geneticallyengineered soluble fusion proteins comprising a polypeptide of thepresent invention, or a fragment thereof, and various portions of theconstant regions of heavy or light chains of immunoglobulins of varioussubclasses (IgG, IgM, IgA, IgE). Preferred as an immunoglobulin is theconstant part of the heavy chain of human IgG, particularly IgGI, wherefusion takes place at the hinge region. In a particular embodiment, theFc part can be removed simply by incorporation of a cleavage sequencewhich can be cleaved with blood clotting factor Xa. Furthermore, thisinvention relates to processes for the preparation of these fusionproteins by genetic engineering, and to the use thereof for drugscreening, diagnosis and therapy. A further aspect of the invention alsorelates to polynucleotides encoding such fusion proteins. Examples offusion protein technology can be found in International PatentApplication Nos. WO94/29458 and WO94/22914.

Another aspect of the invention relates to a method for inducing animmunological response in a mammal which comprises inoculating themammal with a polypeptide of the present invention, adequate to produceantibody and/or T cell immune response to protect said animal from theDiseases hereinbefore mentioned, amongst others. Yet another aspect ofthe invention relates to a method of inducing immunological response ina mammal which comprises, delivering a polypeptide of the presentinvention via a vector directing expression of the polynucleotide andcoding for the polypeptide in vivo in order to induce such animmunological response to produce antibody to protect said animal fromdiseases.

A further aspect of the invention relates to an immunological/vaccineformulation (composition) which, when introduced into a mammalian host,induces an immunological response in that mammal to a polypeptide of thepresent invention wherein the composition comprises a polypeptide orpolynucleotide of the present invention. The vaccine formulation mayfurther comprise a suitable carrier. Since a polypeptide may be brokendown in the stomach, it is preferably administered parenterally (forinstance, subcutaneous, intramuscular, intravenous, or intradermalinjection). Formulations suitable for parenteral administration includeaqueous and non-aqueous sterile injection solutions which may containanti-oxidants, buffers, bacteriostats and solutes which render theformulation instonic with the blood of the recipient; and aqueous andnon-aqueous sterile suspensions which may include suspending agents orthickening agents. The formulations may be presented in unit-dose ormulti-dose containers, for example, sealed ampoules and vials and may bestored in a freeze-dried condition requiring only the addition of thesterile liquid carrier immediately prior to use. The vaccine formulationmay also include adjuvant systems for enhancing the immunogenicity ofthe formulation, such as oil-in water systems and other systems known inthe art. The dosage will depend on the specific activity of the vaccineand can be readily determined by routine experimentation.

Polypeptides of the present invention are responsible for one or morebiological functions, including one or more disease states, inparticular the Diseases hereinbefore mentioned. It is therefore desirousto devise screening methods to identify compounds which stimulate orwhich inhibit the function of the polypeptide. Accordingly, in a furtheraspect, the present invention provides for a method of screeningcompounds to identify those which stimulate or which inhibit thefunction of the polypeptide. In general, agonists or antagonists may beemployed for therapeutic and prophylactic purposes for such Diseases ashereinbefore mentioned. Compounds may be identified from a variety ofsources, for example, cells, cell-free preparations, chemical libraries,and natural product mixtures. Such agonists, antagonists or inhibitorsso-identified may be natural or modified substrates, ligands, receptors,enzymes, etc., as the case may be, of the polypeptide; or may bestructural or functional mimetics thereof (see Coligan et al., CurrentProtocols in Immunology 1(2):Chapter 5 (1991)).

The screening method may simply measure the binding of a candidatecompound to the polypeptide, or to cells or membranes bearing thepolypeptide, or a fusion protein thereof by means of a label directly orindirectly associated with the candidate compound. Alternatively, thescreening method may involve competition with a labeled competitor.Further, these screening methods may test whether the candidate compoundresults in a signal generated by activation or inhibition of thepolypeptide, using detection systems appropriate to the cells bearingthe polypeptide. Inhibitors of activation are generally assayed in thepresence of a known agonist and the effect on activation by the agonistby the presence of the candidate compound is observed. Constitutivelyactive polypeptides may be employed in screening methods for inverseagonists or inhibitors, in the absence of an agonist or inhibitor, bytesting whether the candidate compound results in inhibition ofactivation of the polypeptide. Further, the screening methods may simplycomprise the steps of mixing a candidate compound with a solutioncontaining a polypeptide of the present invention, to form a mixture,measuring VANILREP1 activity in the mixture, and comparing the VANILREP1activity of the mixture to a standard. Fusion proteins, such as thosemade from Fc portion and VANILREP1 polypeptide, as hereinbeforedescribed, can also be used for high-throughput screening assays toidentify antagonists for the polypeptide of the present invention (seeD. Bennett et al., J Mol Recognition, 8:52-58 (1995); and K. Johanson etal., J Biol Chem, 270(16):9459-9471 (1995)).

The polynucleotides, polypeptides and antibodies to the polypeptide ofthe present invention may also be used to configure screening methodsfor detecting the effect of added compounds on the production of mRNAand polypeptide in cells. For example, an ELISA assay may be constructedfor measuring secreted or cell associated levels of polypeptide usingmonoclonal and polyclonal antibodies by standard methods known in theart. This can be used to discover agents which may inhibit or enhancethe production of polypeptide (also called antagonist or agonist,respectively) from suitably manipulated cells or tissues.

The polypeptide may be used to identify membrane bound or solublereceptors, if any, through standard receptor binding techniques known inthe art. These include, but are not limited to, ligand binding andcrosslinking assays in which the polypeptide is labeled with aradioactive isotope (for instance, ¹²⁵I), chemically modified (forinstance, biotinylated), or fused to a peptide sequence suitable fordetection or purification, and incubated with a source of the putativereceptor (cells, cell membranes, cell supernatants, tissue extracts,bodily fluids). Other methods include biophysical techniques such assurface plasmon resonance and spectroscopy. These screening methods mayalso be used to identify agonists and antagonists of the polypeptidewhich compete with the binding of the polypeptide to its receptors, ifany. Standard methods for conducting such assays are well understood inthe art.

Examples of potential polypeptide antagonists include antibodies or, insome cases, oligonucleotides or proteins which are closely related tothe ligands, substrates, receptors, enzymes, etc., as the case may be,of the polypeptide, e.g., a fragment of the ligands, substrates,receptors, enzymes, etc.; or small molecules which bind to thepolypeptide of the present invention but do not elicit a response, sothat the activity of the polypeptide is prevented.

Thus, in another aspect, the present invention relates to a screeningkit for identifying agonists, antagonists, ligands, receptors,substrates, enzymes, etc. for polypeptides of the present invention; orcompounds which decrease or enhance the production of such polypeptides,which comprises:

(a) a polypeptide of the present invention;

(b) a recombinant cell expressing a polypeptide of the presentinvention;

(c) a cell membrane expressing a polypeptide of the present invention;or

(d) antibody to a polypeptide of the present invention;

which polypeptide is preferably that of SEQ ID NO:2 or SEQ ID NO:8.

It will be appreciated that in any such kit, (a), (b), (c) or (d) maycomprise a substantial component.

It will be readily appreciated by the skilled artisan that a polypeptideof the present invention may also be used in a method for thestructure-based design of an agonist, antagonist or inhibitor of thepolypeptide, by:

(a) determining in the first instance the three-dimensional structure ofthe polypeptide;

(b) deducing the three-dimensional structure for the likely reactive orbinding site(s) of an agonist, antagonist or inhibitor;

(c) synthesing candidate compounds that are predicted to bind to orreact with the deduced binding or reactive site; and

(d) testing whether the candidate compounds are indeed agonists,antagonists or inhibitors.

It will be further appreciated that this will normally be an iterativeprocess.

In a further aspect, the present invention provides methods of treatingabnormal conditions such as, for instance, pain, chronic pain,neuropathic pain, postoperative pain, rheumatoid arthritic pain,neuralgia, neuropathies, algesia, nerve injury, ischaemia,neurodegeneration, stroke, incontinence and inflammatory disorders,related to either an excess of, or an under-expression of, VANILREP1polypeptide activity.

If the activity of the polypeptide is in excess, several approaches areavailable. One approach comprises administering to a subject in needthereof an inhibitor compound (antagonist) as hereinabove described,optionally in combination with a pharmaceutically acceptable carrier, inan amount effective to inhibit the function of the polypeptide, such as,for example, by blocking the binding of ligands, substrates, receptors,enzymes, etc., or by inhibiting a second signal, and thereby alleviatingthe abnormal condition. In another approach, soluble forms of thepolypeptides still capable of binding the ligand, substrate, enzymes,receptors, etc. in competition with endogenous polypeptide may beadministered. Typical examples of such competitors include fragments ofthe VANILREP1 polypeptide.

In still another approach, expression of the gene encoding endogenousVANILREP1 polypeptide can be inhibited using expression blockingtechniques. Known such techniques involve the use of antisensesequences, either internally generated or externally administered (see,for example, O'Connor, J Neurochem (1991) 56:560 inOligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRCPress, Boca Raton, Fla. (1988)). Alternatively, oligonucleotides whichform triple helices (“triplexes”) with the gene can be supplied (see,for example, Lee et al., Nucleic Acids Res (1979) 6:3073; Cooney et al.,Science (1988) 241:456; Dervan et al., Science (1991) 251:1360). Theseoligomers can be administeredper se or the relevant oligomers can beexpressed in vivo. Synthetic antisense or triplex oligonucleotides maycomprise modified bases or modified backbones. Examples of the latterinclude methylphosphonate, phosphorothioate or peptide nucleic acidbackbones. Such backbones are incorporated in the antisense or triplexoligonucleotide in order to provide protection from degradation bynucleases and are well known in the art. Antisense and triplex moleculessynthesised with these or other modified backbones also form part of thepresent invention.

In addition, expression of the human VANILREP1 polypeptide may beprevented by using ribozymes specific to the human VANILREP1 mRNAsequence. Ribozymes are catalytically active RNAs that can be natural orsynthetic (see for example Usman, N, et al., Curr. Opin. Struct. Biol(1996) 6(4), 527-33.) Synthetic ribozymes can be designed tospecifically cleave human VANILREP1 mRNAs at selected positions therebypreventing translation of the human VANILREP1 mRNAs into functionalpolypeptide. Ribozymes may be synthesised with a natural ribosephosphate backbone and natural bases, as normally found in RNAmolecules. Alternatively the ribosymes may be synthesised withnon-natural backbones to provide protection from ribonucleasedegradation, for example, 2′-O-methyl RNA, and may contain modifiedbases.

For treating abnormal conditions related to an under-expression ofVANILREP1 and its activity, several approaches are also available. Oneapproach comprises administering to a subject a therapeuticallyeffective amount of a compound which activates a polypeptide of thepresent invention, i.e., an agonist as described above, in combinationwith a pharmaceutically acceptable carrier, to thereby alleviate theabnormal condition. Alternatively, gene therapy may be employed toeffect the endogenous production of VANILREP1 by the relevant cells inthe subject. For example, a polynucleotide of the invention may beengineered for expression in a replication defective retroviral vector,as discussed above. The retroviral expression construct may then beisolated and introduced into a packaging cell transduced with aretroviral plasmid vector containing RNA encoding a polypeptide of thepresent invention such that the packaging cell now produces infectiousviral particles containing the gene of interest. These producer cellsmay be administered to a subject for engineering cells in vivo andexpression of the polypeptide in vivo. For an overview of gene therapy,see Chapter 20, Gene Therapy and other Molecular Genetic-basedTherapeutic Approaches, (and references cited therein) in HumanMolecular Genetics, T Strachan and A P Read, BIOS Scientific PublishersLtd (1996). Another approach is to administer a therapeutic amount of apolypeptide of the present invention in combination with a suitablepharmaceutical carrier.

In a further aspect, the present invention provides for pharmaceuticalcompositions comprising a therapeutically effective amount of apolypeptide, such as the soluble form of a polypeptide of the presentinvention, agonist/antagonist peptide or small molecule compound, incombination with a pharmaceutically acceptable carrier or excipient.Such carriers include, but are not limited to, saline, buffered saline,dextrose, water, glycerol, ethanol, and combinations thereof Theinvention further relates to pharmaceutical packs and kits comprisingone or more containers filled with one or more of the ingredients of theaforementioned compositions of the invention. Polypeptides and othercompounds of the present invention may be employed alone or inconjunction with other compounds, such as therapeutic compounds.

The composition will be adapted to the route of administration, forinstance by a systemic or an oral route. Preferred forms of systemicadministration include injection, typically by intravenous injection.Other injection routes, such as subcutaneous, intramuscular, orintraperitoneal, can be used. Alternative means for systemicadministration include transmucosal and transdermal administration usingpenetrants such as bile salts or fusidic acids or other detergents. Inaddition, if a polypeptide or other compounds of the present inventioncan be formulated in an enteric or an encapsulated formulation, oraladministration may also be possible. Administration of these compoundsmay also be topical and/or localized, in the form of salves, pastes,gels, and the like.

The dosage range required depends on the choice of peptide or othercompounds of the present invention, the route of administration, thenature of the formulation, the nature of the subject's condition, andthe judgment of the attending practitioner. Suitable dosages, however,are in the range of 0.1-100 μg/kg of subject. Wide variations in theneeded dosage, however, are to be expected in view of the variety ofcompounds available and the differing efficiencies of various routes ofadministration. For example, oral administration would be expected torequire higher dosages than administration by intravenous injection.Variations in these dosage levels can be adjusted using standardempirical routines for optimization, as is well understood in the art.

Polypeptides used in treatment can also be generated endogenously in thesubject, in treatment modalities often referred to as “gene therapy” asdescribed above. Thus, for example, cells from a subject may beengineered with a polynucleotide, such as a DNA or RNA, to encode apolypeptide ex Vivo, and for example, by the use of a retroviral plasmidvector. The cells are then introduced into the subject.

Polynucleotide and polypeptide sequences form a valuable informationresource with which to identify further sequences of similar homology.This is most easily facilitated by storing the sequence in a computerreadable medium and then using the stored data to search a sequencedatabase using well known searching tools, such as those in the GCG andLasergene software packages. Accordingly, in a further aspect, thepresent invention provides for a computer readable medium having storedthereon a polynucleotide comprising the sequence of SEQ ID NO:1 or SEQID NO:7 and/or a polypeptide sequence encoded thereby.

The following definitions are provided to facilitate understanding ofcertain terms used frequently hereinbefore.

“Antibodies” as used herein includes polyclonal and monoclonalantibodies, chimeric, single chain, and humanized antibodies, as well asFab fragments, including the products of an Fab or other immunoglobulinexpression library.

“Isolated” means altered “by the hand of man” from the natural state. Ifan “isolated” composition or substance occurs in nature, it has beenchanged or removed from its original environment, or both. For example,a polynucleotide or a polypeptide naturally present in a living animalis not “isolated,” but the same polynucleotide or polypeptide separatedfrom the coexisting materials of its natural state is “isolated”, as theterm is employed herein.

“Polynucleotide” generally refers to any polyribonucleotide orpolydeoxribonucleotide, which may be unmodified RNA or DNA or modifiedRNA or DNA. “Polynucleotides” include, without limitation, single- anddouble-stranded DNA, DNA that is a mixture of single- anddouble-stranded regions, single- and double-stranded RNA, and RNA thatis mixture of single- and double-stranded regions, hybrid moleculescomprising DNA and RNA that may be single-stranded or, more typically,double-stranded or a mixture of single- and double-stranded regions. Inaddition, “polynucleotide” refers to triple-stranded regions comprisingRNA or DNA or both RNA and DNA. The term “polynucleotide” also includesDNAs or RNAs containing one or more modified bases and DNAs or RNAs withbackbones modified for stability or for other reasons. “Modified” basesinclude, for example, tritylated bases and unusual bases such asinosine. A ariety of modifications may be made to DNA and RNA; thus,“polynucleotide” embraces chemically, enzymatically or metabolicallymodified forms of polynucleotides as typically found in nature, as wellas the chemical forms of DNA and RNA characteristic of viruses andcells. “Polynucleotide” also embraces relatively short polynucleotides,often referred to as oligonucleotides.

“Polypeptide” refers to any peptide or protein comprising two or moreamino acids joined to each other by peptide bonds or modified peptidebonds, i.e., peptide isosteres. “Polypeptide” refers to both shortchains, commonly referred to as peptides, oligopeptides or oligomers,and to longer chains, generally referred to as proteins. Polypeptidesmay contain amino acids other than the 20 gene-encoded amino acids.“Polypeptides” include amino acid sequences modified either by naturalprocesses, such as post-translational processing, or by chemicalmodification techniques which are well known in the art. Suchmodifications are well described in basic texts and in more detailedmonographs, as well as in a voluminous research literature.Modifications may occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxyl termini.It will be appreciated that the same type of modification may be presentto the same or varying degrees at several sites in a given polypeptide.Also, a given polypeptide may contain many types of modifications.Polypeptides may be branched as a result of ubiquitination, and they maybe cyclic, with or without branching. Cyclic, branched and branchedcyclic polypeptides may result from post-translation natural processesor may be made by synthetic methods. Modifications include acetylation,acylation, ADP-ribosylation, amidation, biotinylation, covalentattachment of flavin, covalent attachment of a heme moiety, covalentattachment of a nucleotide or nucleotide derivative, covalent attachmentof a lipid or lipid derivative, covalent attachment ofphosphotidylinositol, cross-linking, cyclization, disulfide bondformation, demethylation, formation of covalent cross-links, formationof cystine, formation of pyroglutamate, formylation,gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation,iodination, methylation, myristoylation, oxidation, proteolyticprocessing, phosphorylation, prenylation, racemization, selenoylation,sulfation, transfer-RNA mediated addition of amino acids to proteinssuch as arginylation, and ubiquitination (see, for instance,Proteins—Structure and Molecular Properties, 2nd Ed., T. E. Creighton,W. H. Freeman and Company, New York, 1993; Wold, F., Post-translationalProtein Modifications: Perspectives and Prospects, pgs. 1-12 inPost-translational Covalent Modification of Proteins, B. C. Johnson,Ed., Academic Press, New York, 1983; Seifter et al., “Analysis forprotein modifications and nonprotein cofactors”, Meth Enzymol (1990)182:626-646 and Rattan et al., “Protein Synthesis: Post-translationalModifications and Aging”, Ann NY Acad Sci (1992) 663:48-62).

“Variant” refers to a polynucleotide or polypeptide that differs from areference polynucleotide or polypeptide, but retains essentialproperties. A typical variant of a polynucleotide differs in nucleotidesequence from another, reference polynucleotide. Changes in thenucleotide sequence of the variant may or may not alter the amino acidsequence of a polypeptide encoded by the reference polynucleotide.Nucleotide changes may result in amino acid substitutions, additions,deletions, fusions and truncations in the polypeptide encoded by thereference sequence, as discussed below. A typical variant of apolypeptide differs in amino acid sequence from another, referencepolypeptide. Generally, differences are limited so that the sequences ofthe reference polypeptide and the variant are closely similar overalland, in many regions, identical. A variant and reference polypeptide maydiffer in amino acid sequence by one or more substitutions, additions,deletions in any combination. A substituted or inserted amino acidresidue may or may not be one encoded by the genetic code. A variant ofa polynucleotide or polypeptide may be a naturally occurring such as anallelic variant, or it may be a variant that is not known to occurnaturally. Non-naturally occurring variants of polynucleotides andpolypeptides may be made by mutagenesis techniques or by directsynthesis.

“Identity,” as known in the art, is a relationship between two or morepolypeptide sequences or two or more polynucleotide sequences, asdetermined by comparing the sequences. In the art, “identity” also meansthe degree of sequence relatedness between polypeptide or polynucleotidesequences, as the case may be, as determined by the match betweenstrings of such sequences. “Identity” and “similarity” can be readilycalculated by known methods, including but not limited to thosedescribed in (Computational Molecular Biology, Lesk, A. M., ed., OxfordUniversity Press, New York, 1988; Biocomputing: Informatics and GenomeProjects, Smith, D. W., ed., Academic Press, New York, 1993; ComputerAnalysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G.,eds., Humana Press, New Jersey, 1994; Sequence Analysis in MolecularBiology, von Heinje, G., Academic Press, 1987; and Sequence AnalysisPrimer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York,1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073(1988). Preferred methods to determine identity are designed to give thelargest match between the sequences tested. Methods to determineidentity and similarity are codified in publicly available computerprograms. Preferred computer program methods to determine identity andsimilarity between two sequences include, but are not limited to, theGCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387 (1984)), BLASTP, BLASTN, and FASTA (Atschul, S. F. et al., J. Molec.Biol. 215: 403-410 (1990). The BLAST X program is publicly availablefrom NCBI and other sources (BLAS TManual, Altschul, S., et al., NCBINLM NIH Bethesda, MD 20894; Altschul, S., et al., J. Mol Biol. 215:403-410 (1990). The well known Smith Waterman algorithm may also be usedto determine identity.

Preferred parameters for polypeptide sequence comparison include thefollowing:

1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970)

Comparison matrix: BLOSSUM62 from Hentikoff and Hentikoff, Proc. Natl.Acad. Sci. USA. 89:10915-10919 (1992)

Gap Penalty: 12

Gap Length Penalty: 4

A program useful with these parameters is publicly available as the“gap” program from Genetics Computer Group, Madison Wis. Theaforementioned parameters are the default parameters for peptidecomparisons (along with no penalty for end gaps).

Preferred parameters for polynucleotide comparison include thefollowing:

1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970)

Comparison matrix: matches=+10, mismatch=0

Gap Penalty: 50

Gap Length Penalty: 3

Available as: The “gap” program from Genetics Computer Group, MadisonWI. These are the default parameters for nucleic acid comparisons.

By way of example, a polynucleotide sequence of the present inventionmay be identical to the reference sequence of SEQ ID NO:1, that is be100% identical, or it may include up to a certain integer number ofnucleotide alterations as compared to the reference sequence. Suchalterations are selected from the group consisting of at least onenucleotide deletion, substitution, including transition andtransversion, or insertion, and wherein said alterations may occur atthe 5′ or 3′ terminal positions of the reference nucleotide sequence oranywhere between those terminal positions, interspersed eitherindividually among the nucleotides in the reference sequence or in oneor more contiguous groups within the reference sequence. The number ofnucleotide alterations is determined by multiplying the total number ofnucleotides in SEQ ID NO:1 by the numerical percent of the respectivepercent identity(divided by 100) and subtracting that product from saidtotal number of nucleotides in SEQ ID NO:1, or:

n _(n) ≦x _(n)−(x _(n) ·y)

wherein n_(n) is the number of nucleotide alterations, x_(n) is thetotal number of nucleotides in SEQ ID NO:1, and y is, for instance, 0.70for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95%, etc.,and wherein any non-integer product of x_(n) and y is rounded down tothe nearest integer prior to subtracting it from x_(n). Alterations of apolynucleotide sequence encoding the polypeptide of SEQ ID NO:2 maycreate nonsense, missense or frameshift mutations in this codingsequence and thereby alter the polypeptide encoded by the polynucleotidefollowing such alterations.

Similarly, a polypeptide sequence of the present invention may beidentical to the reference sequence of SEQ ID NO:2, that is be 100%identical, or it may include up to a certain integer number of aminoacid alterations as compared to the reference sequence such that the %identity is less than 100%. Such alterations are selected from the groupconsisting of at least one amino acid deletion, substitution, includingconservative and non-conservative substitution, or insertion, andwherein said alterations may occur at the amino- or carboxy-terminalpositions of the reference polypeptide sequence or anywhere betweenthose terminal positions, interspersed either individually among theamino acids in the reference sequence or in one or more contiguousgroups within the reference sequence. The number of amino acidalterations for a given % identity is determined by multiplying thetotal number of amino acids in SEQ ID NO:2 by the numerical percent ofthe respective percent identity(divided by 100) and then subtractingthat product from said total number of amino acids in SEQ ID NO:2, or:

n _(a) ≦x _(a)−(xa·y),

wherein n_(a) is the number of amino acid alterations, x_(a) is thetotal number of amino acids in SEQ ID NO:2, and y is, for instance 0.70for 70%, 0.80 for 80%, 0.85 for 85% etc., and wherein any non-integerproduct of x_(a) and y is rounded down to the nearest integer prior tosubtracting it from x_(a).

“Homolog” is a generic term used in the art to indicate a polynucleotideor polypeptide sequence possessing a high degree of sequence relatednessto a subject sequence. Such relatedness may be quantified by determiningthe degree of identity and/or similarity between the sequences beingcompared as hereinbefore described. Falling within this generic term arethe terms “ortholog”, meaning a polynucleotide or polypeptide that isthe functional equivalent of a polynucleotide or polypeptide in anotherspecies, and “paralog” meaning a functionally similar sequence whenconsidered within the same species.

“Fusion protein” refers to a protein encoded by two, often unrelated,fused genes or fragments thereof In one example, EP-A-0 464 disclosesfusion proteins comprising various portions of constant region ofimmunoglobulin molecules together with another human protein or partthereof. In many cases, employing an immunoglobulin Fc region as a partof a fusion protein is advantageous for use in therapy and diagnosisresulting in, for example, improved pharmacokinetic properties [see,e.g., EP-A 0232 262]. On the other hand, for some uses it would bedesirable to be able to delete the Fc part after the fusion protein hasbeen expressed, detected and purified.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

EXAMPLE Example 1 Electrophysiological Studies

Xenopus laevis oocyte removal and dissociation were performed.Injections of cDNA for VANILREP1 were made into the nuclei ofdefolliculated oocytes (1.5 ng cDNA/oocyte). After injection the oocyteswere incubated between 19-22° C. in modified Barth's solution (MBS) plusgentamycin (0.1 mg/ml, pH 7.4) and used for electrophysiologicalrecordings within 24 days.

For electrophysiological recordings oocytes were placed in a recordingchamber and continuously perfused with a solution containing in mM: NaCl88, KCl 1, NaHCO₃ 2.4, HEPES 15, MgCl₂ 1, BaCl₂ 0.1. (14 ml min⁻¹).Solution was applied using large bore tubing (internal diameter 1.5 mm)which facilitated rapid solution exchange (half-time 350-1000 ms).Oocytes were held under voltage clamp at −60 mV using the two-electrodevoltage-clamp technique. Electrodes were low resistance (0.5-3 MΩ) andwere filled with 3 M KCl.

Currents were evoked in response to application of capsaicin appliedthrough the perfusion system until the maximum current amplitude wasreached. FIG. 1 shows currents evoked from a cell in response tocapsaicin over a range of concentrations. FIG. 2 shows inhibition of thecapsaicin evoked response (1 ) by capsazepine (10 uM). The inhibition isreversible with extended washing.

8 1 4803 DNA HOMO SAPIENS 1 cagcgtcggg tgcagtttgg ccggaggttg cagtgagcagagattgcccc attgcactct 60 agtctgggcg acagggtgag acacacacac acagacacacacacacacac acacacacac 120 acacaagcct aaacattcra ggccaggatg cttgacagatgttgattcat aaaaatgaca 180 aaaagcacaa aatccaaaat ctcgtataag ctcagtggctgtggcagcga ggttgaagag 240 caaaggcagg ccgggcacct ggctgatgat gtgtggacccgttgcacagc agggccccgc 300 agtgcggtgt gggtgtgggt gtgggtgggc cagtytctgccgctcaccct attccaggga 360 cacagtctgc ttggctcttc tggactgagc catcctcatcaccgagatcc tccctgaatt 420 cagcccacga cagccacccc ggccgttttc cttgttctgtgtggggaggg aggcagcgcg 480 gtggttatca acctcaccct gcagaggagg cacctgaggcccagagacga ggagggatgg 540 gtctaaccca gaaccacaga tggctctgag ccgggggcctgtccaccctc ccaggccgac 600 gtcagtggcc gcaggactgc ctgggccctg ctaggcctgctcacctctga ggcctctggg 660 gtgagaggtt cagtcctgga aacacttcag ttctagggggctgggggcag cagcaagttg 720 gagttttggg gtaccctgct tcacagggcc cttggcaaggagggcaggtg gggtctaagg 780 acaagcagtc cttactttgg gagtcaaccc cggcgtggtggctgctgcag gttgcacact 840 gggccacaga ggatccagca aggatgaaga aatggagcagcacagacttg ggggcagctg 900 cggacccact ccaaaaggac acctgcccag accccctggatggagaccct aactccaggc 960 cacctccagc caagccccag ctctccacgg ccaagagccgcacccggctc tttgggaagg 1020 gtgactcgga ggaggctttc ccggtggatt gccctcacgaggaaggtgag ctggactcct 1080 gcccgaccat cacagtcagc cctgttatca ccatccagaggccaggagac ggccccaccg 1140 gtgccaggct gctgtcccag gactctgtcg ccgccagcaccgagaagacc ctcaggctct 1200 atgatcgcag gagtatcttt gaagccgttg ctcagaataactgccaggat ctggagagcc 1260 tgctgctctt cctgcagaag agcaagaagc acctcacagacaacgagttc aaagaccctg 1320 agacagggaa gacctgtctg ctgaaagcca tgctcaacctgcacgacgga cagaacacca 1380 ccatccccct gctcctggag atcgcgcggc aaacggacagcctgaaggag cttgtcaacg 1440 ccagctacac ggacagctac tacaagggcc agacagcactgcacatcgcc atcgagagac 1500 gcaacatggc cctggtgacc ctcctggtgg agaacggagcagacgtccag gctgcggccc 1560 atggggactt ctttaagaaa accaaagggc ggcctggattctacttcggt gaactgcccc 1620 tgtccctggc cgcgtgcacc aaccagctgg gcatcgtgaagttcctgctg cagaactcct 1680 ggcagacggc cgacatcagc gccagggact cggtgggcaacacggtgctg cacgccctgg 1740 tggaggtggc cgacaacacg gccgacaaca cgaagtttgtgacgagcatg tacaatgaga 1800 ttctgatcct gggggccaaa ctgcacccga cgctgaagctggaggagctc accaacaaga 1860 agggaatgac gccgctggct ctggcagctg ggaccgggaagatcggggtc ttggcctata 1920 ttctccagcg ggagatccag gagcccgagt gcaggcacctgtccaggaag ttcaccgagt 1980 gggcctacgg gcccgtgcac tcctcgctgt acgacctgtcctgcatcgac acctgcgaga 2040 agaactcggt gctggaggtg atcgcctaca gcagcagcgagacccctaat cgccacgaca 2100 tgctcttggt ggagccgctg aaccgactcc tgcaggacaagtgggacaga ttcgtcaagc 2160 gcatcttcta cttcaacttc ctggtctact gcctgtacatgatcatcttc accatggctg 2220 cctactacag gcccgtggat ggcttgcctc cctttaagatggaaaaaact ggagactatt 2280 tccgagttac tggagagatc ctgtctgtgt taggaggagtctacttcttt ttccgaggga 2340 ttcagtattt cctgcagagg cggccgtcga tgaagaccctgtttgtggac agctacagtg 2400 agatgctttt ctttctgcag tcactgttca tgctggccaccgtggtgctg tacttcagcc 2460 acctcaagga gtatgtggct tccatggtat tctccctggccttgggctgg accaacatgc 2520 tctactacac ccgcggtttc cagcagatgg gcatctatgccgtcatgata gagaagatga 2580 tcctgagaga cctgtgccgt ttcatgtttg tctacatcgtcttcttgttc gggttttcca 2640 cagcggtggt gacgctgatt gaagacggga agaatgactccctgccgtct gagtccacgt 2700 cgcacaggtg gcgggggcct gcctgcaggc cccccgatagctcctacaac agcctgtact 2760 ccacctgcct ggagctgttc aagttcacca tcggcatgggcgacctggag ttcactgaga 2820 actatgactt caaggctgtc ttcatcatcc tgctgctggcctatgtaatt ctcacctaca 2880 tcctcctgct caacatgctc atcgccctca tgggtgagactgtcaacaag atcgcacagg 2940 agagcaagaa catctggaag ctgcagagag ccatcaccatcctggacacg gagaagagct 3000 tccttaagtg catgaggaag gccttccgct caggcaagctgctgcaggtg gggtacacac 3060 ctgatggcaa ggacgactac cggtggtgct tcagggtggacgaggtgaac tggaccacct 3120 ggaacaccaa cgtgggcatc atcaacgaag acccgggcaactgtgagggc gtcaagcgca 3180 ccctgagctt ctccctgcgg tcaagcagag tttcaggcagacactggaag aactttgccc 3240 tggtccccct tttaagagag gcaagtgctc gagataggcagtctgctcag cccgaggaag 3300 tttatctgcg acagttttca gggtctctga agccagaggacgctgaggtc ttcaagagtc 3360 ctgccgcttc cggggagaag tgaggacgtc acgcagacagcactgtcaac actgggcctt 3420 aggagacccc gttgccacgg ggggctgctg agggaacaccagtgctctgt cagcagcctg 3480 gcctggtctg tgcctgccca gcatgttccc aaatctgtgctggacaagct gtgggaagcg 3540 ttcttggaag catggggagt gatgtacatc caaccgtcactgtccccaag tgaatctcct 3600 aacagacttt caggttttta ctcactttac taaacagtktggatggtcag tctctactgg 3660 gacatgttag gcccttgttt tctttgattt tattcttttttttgagacag aatttcactc 3720 ttctcaccca ggctggaatg cagtggcaca attttggctccctgcaacct ccgcctcctg 3780 gattccagca attctcctgc ctcggcttcc caagtagctgggattacagg cacgtgccac 3840 catgtctggc taattttttg tattttttta atagatatggggtttcgcca tgttggccag 3900 gctggtctcg aactcctgac ctcaggtgat ccgcccacctcggcctccca aagtgctggg 3960 attacaggtg tgagcctcca cacctggctg ttttctttgattttattctt tttttttttt 4020 tctgtgagac agagtttcac tcttgttgcc caggctggagtgcagtggtg tgatcttggc 4080 tcactgcaac ttctgcctcc cgggttcaag cgattcttctgcttcagtct cccaagtagc 4140 ttggattaca ggtgagcact accacgcccg gctaatttttgtatttttaa taragacggg 4200 gtttcaccat gttggccagg ctggtctcga actcttgacctcaggtgatc tgcccgcctt 4260 ggcctcccaa agtgctggga ttacaggtgt gagccgctgcgctcggcctt ctttgatttt 4320 atattattag gagcaaaagt aaatgaagcc caggaaaacacctttgggaa caaactcttc 4380 ctttgatgga aaatgcagag gcccttcctc tctgtgccgtgcttgctcct cttacctgcc 4440 cgggtggttt gggggtgttg gtgtttcctc cctggagaagatgggggagg ctgtcccact 4500 cccagctctg gcagaatcaa gctgttgcag cagtgccttcttcatccttc cttacgatca 4560 atcacagtct ccagaagatc agctcaattg ctgtgcaggttaaaactaca gaaccacatc 4620 ccaaaggtac ctggtaagaa tgtttgaaag atcttccatttctaggaacc ccagtcctgc 4680 ttctccgcaa tggcacatgc ttccactcca tccatactggcatcctcaaa taaacagata 4740 tgtatacaat aaaaaaaaaa aaaaaaaaaa rrgcggccgctgaattctag acctgcccgg 4800 gcg 4803 2 839 PRT HOMO SAPIENS 2 Met Lys LysTrp Ser Ser Thr Asp Leu Gly Ala Ala Ala Asp Pro Leu 1 5 10 15 Gln LysAsp Thr Cys Pro Asp Pro Leu Asp Gly Asp Pro Asn Ser Arg 20 25 30 Pro ProPro Ala Lys Pro Gln Leu Ser Thr Ala Lys Ser Arg Thr Arg 35 40 45 Leu PheGly Lys Gly Asp Ser Glu Glu Ala Phe Pro Val Asp Cys Pro 50 55 60 His GluGlu Gly Glu Leu Asp Ser Cys Pro Thr Ile Thr Val Ser Pro 65 70 75 80 ValIle Thr Ile Gln Arg Pro Gly Asp Gly Pro Thr Gly Ala Arg Leu 85 90 95 LeuSer Gln Asp Ser Val Ala Ala Ser Thr Glu Lys Thr Leu Arg Leu 100 105 110Tyr Asp Arg Arg Ser Ile Phe Glu Ala Val Ala Gln Asn Asn Cys Gln 115 120125 Asp Leu Glu Ser Leu Leu Leu Phe Leu Gln Lys Ser Lys Lys His Leu 130135 140 Thr Asp Asn Glu Phe Lys Asp Pro Glu Thr Gly Lys Thr Cys Leu Leu145 150 155 160 Lys Ala Met Leu Asn Leu His Asp Gly Gln Asn Thr Thr IlePro Leu 165 170 175 Leu Leu Glu Ile Ala Arg Gln Thr Asp Ser Leu Lys GluLeu Val Asn 180 185 190 Ala Ser Tyr Thr Asp Ser Tyr Tyr Lys Gly Gln ThrAla Leu His Ile 195 200 205 Ala Ile Glu Arg Arg Asn Met Ala Leu Val ThrLeu Leu Val Glu Asn 210 215 220 Gly Ala Asp Val Gln Ala Ala Ala His GlyAsp Phe Phe Lys Lys Thr 225 230 235 240 Lys Gly Arg Pro Gly Phe Tyr PheGly Glu Leu Pro Leu Ser Leu Ala 245 250 255 Ala Cys Thr Asn Gln Leu GlyIle Val Lys Phe Leu Leu Gln Asn Ser 260 265 270 Trp Gln Thr Ala Asp IleSer Ala Arg Asp Ser Val Gly Asn Thr Val 275 280 285 Leu His Ala Leu ValGlu Val Ala Asp Asn Thr Ala Asp Asn Thr Lys 290 295 300 Phe Val Thr SerMet Tyr Asn Glu Ile Leu Ile Leu Gly Ala Lys Leu 305 310 315 320 His ProThr Leu Lys Leu Glu Glu Leu Thr Asn Lys Lys Gly Met Thr 325 330 335 ProLeu Ala Leu Ala Ala Gly Thr Gly Lys Ile Gly Val Leu Ala Tyr 340 345 350Ile Leu Gln Arg Glu Ile Gln Glu Pro Glu Cys Arg His Leu Ser Arg 355 360365 Lys Phe Thr Glu Trp Ala Tyr Gly Pro Val His Ser Ser Leu Tyr Asp 370375 380 Leu Ser Cys Ile Asp Thr Cys Glu Lys Asn Ser Val Leu Glu Val Ile385 390 395 400 Ala Tyr Ser Ser Ser Glu Thr Pro Asn Arg His Asp Met LeuLeu Val 405 410 415 Glu Pro Leu Asn Arg Leu Leu Gln Asp Lys Trp Asp ArgPhe Val Lys 420 425 430 Arg Ile Phe Tyr Phe Asn Phe Leu Val Tyr Cys LeuTyr Met Ile Ile 435 440 445 Phe Thr Met Ala Ala Tyr Tyr Arg Pro Val AspGly Leu Pro Pro Phe 450 455 460 Lys Met Glu Lys Thr Gly Asp Tyr Phe ArgVal Thr Gly Glu Ile Leu 465 470 475 480 Ser Val Leu Gly Gly Val Tyr PhePhe Phe Arg Gly Ile Gln Tyr Phe 485 490 495 Leu Gln Arg Arg Pro Ser MetLys Thr Leu Phe Val Asp Ser Tyr Ser 500 505 510 Glu Met Leu Phe Phe LeuGln Ser Leu Phe Met Leu Ala Thr Val Val 515 520 525 Leu Tyr Phe Ser HisLeu Lys Glu Tyr Val Ala Ser Met Val Phe Ser 530 535 540 Leu Ala Leu GlyTrp Thr Asn Met Leu Tyr Tyr Thr Arg Gly Phe Gln 545 550 555 560 Gln MetGly Ile Tyr Ala Val Met Ile Glu Lys Met Ile Leu Arg Asp 565 570 575 LeuCys Arg Phe Met Phe Val Tyr Ile Val Phe Leu Phe Gly Phe Ser 580 585 590Thr Ala Val Val Thr Leu Ile Glu Asp Gly Lys Asn Asp Ser Leu Pro 595 600605 Ser Glu Ser Thr Ser His Arg Trp Arg Gly Pro Ala Cys Arg Pro Pro 610615 620 Asp Ser Ser Tyr Asn Ser Leu Tyr Ser Thr Cys Leu Glu Leu Phe Lys625 630 635 640 Phe Thr Ile Gly Met Gly Asp Leu Glu Phe Thr Glu Asn TyrAsp Phe 645 650 655 Lys Ala Val Phe Ile Ile Leu Leu Leu Ala Tyr Val IleLeu Thr Tyr 660 665 670 Ile Leu Leu Leu Asn Met Leu Ile Ala Leu Met GlyGlu Thr Val Asn 675 680 685 Lys Ile Ala Gln Glu Ser Lys Asn Ile Trp LysLeu Gln Arg Ala Ile 690 695 700 Thr Ile Leu Asp Thr Glu Lys Ser Phe LeuLys Cys Met Arg Lys Ala 705 710 715 720 Phe Arg Ser Gly Lys Leu Leu GlnVal Gly Tyr Thr Pro Asp Gly Lys 725 730 735 Asp Asp Tyr Arg Trp Cys PheArg Val Asp Glu Val Asn Trp Thr Thr 740 745 750 Trp Asn Thr Asn Val GlyIle Ile Asn Glu Asp Pro Gly Asn Cys Glu 755 760 765 Gly Val Lys Arg ThrLeu Ser Phe Ser Leu Arg Ser Ser Arg Val Ser 770 775 780 Gly Arg His TrpLys Asn Phe Ala Leu Val Pro Leu Leu Arg Glu Ala 785 790 795 800 Ser AlaArg Asp Arg Gln Ser Ala Gln Pro Glu Glu Val Tyr Leu Arg 805 810 815 GlnPhe Ser Gly Ser Leu Lys Pro Glu Asp Ala Glu Val Phe Lys Ser 820 825 830Pro Ala Ala Ser Gly Glu Lys 835 3 4803 DNA HOMO SAPIENS 3 cagcgtcgggtgcagtttgg ccggaggttg cagtgagcag agattgcccc attgcactct 60 agtctgggcgacagggtgag acacacacac acagacacac acacacacac acacacacac 120 acacaagcctaaacattcra ggccaggatg cttgacagat gttgattcat aaaaatgaca 180 aaaagcacaaaatccaaaat ctcgtataag ctcagtggct gtggcagcga ggttgaagag 240 caaaggcaggccgggcacct ggctgatgat gtgtggaccc gttgcacagc agggccccgc 300 agtgcggtgtgggtgtgggt gtgggtgggc cagtytctgc cgctcaccct attccaggga 360 cacagtctgcttggctcttc tggactgagc catcctcatc accgagatcc tccctgaatt 420 cagcccacgacagccacccc ggccgttttc cttgttctgt gtggggaggg aggcagcgcg 480 gtggttatcaacctcaccct gcagaggagg cacctgaggc ccagagacga ggagggatgg 540 gtctaacccagaaccacaga tggctctgag ccgggggcct gtccaccctc ccaggccgac 600 gtcagtggccgcaggactgc ctgggccctg ctaggcctgc tcacctctga ggcctctggg 660 gtgagaggttcagtcctgga aacacttcag ttctaggggg ctgggggcag cagcaagttg 720 gagttttggggtaccctgct tcacagggcc cttggcaagg agggcaggtg gggtctaagg 780 acaagcagtccttactttgg gagtcaaccc cggcgtggtg gctgctgcag gttgcacact 840 gggccacagaggatccagca aggatgaaga aatggagcag cacagacttg ggggcagctg 900 cggacccactccaaaaggac acctgcccag accccctgga tggagaccct aactccaggc 960 cacctccagccaagccccag ctctccacgg ccaagagccg cacccggctc tttgggaagg 1020 gtgactcggaggaggctttc ccggtggatt gccctcacga ggaaggtgag ctggactcct 1080 gcccgaccatcacagtcagc cctgttatca ccatccagag gccaggagac ggccccaccg 1140 gtgccaggctgctgtcccag gactctgtcg ccgccagcac cgagaagacc ctcaggctct 1200 atgatcgcaggagtatcttt gaagccgttg ctcagaataa ctgccaggat ctggagagcc 1260 tgctgctcttcctgcagaag agcaagaagc acytcacaga caacgagttc aaagaccctg 1320 agacagggaagacctgtctg ctgaaagcca tgctcaacct gcacgacgga cagaacacca 1380 ccatccccctgctcctggag atcgcgcggc aaacggacag cctgaaggag cttgtcaacg 1440 ccrgctacacggacagstac tacaagggcc agacagcact gcacatcgcc atcgagagac 1500 gcaacatggccctggtgacc ctcctggtgg agaacggagc agacgtccag gctgcggccc 1560 atggggacttctttaagaaa accaaagggc ggcctggatt ctacttcggt gaactgcccc 1620 tgtccctggccgcgtgcacc aaccagctgg gcatcgtgaa gttcctgctg cagaactcct 1680 ggcagacggccgacatcagc gccagggact cggtgggcaa cacggtgctg cacgccctgg 1740 tggaggtggccgacaacacg gccgacaaca cgaagtttgt gacgagcatg tacaatgaga 1800 ttctgatcctgggggccaaa ctgcacccga cgctgaagct ggaggagctc accaacaaga 1860 agggaatgacgccgctggct ctggcagctg ggaccgggaa gatcggggtc ttggcctata 1920 ttctccagcgggagatccag gagcccgagt gcaggcacct gtccaggaag ttcaccgagt 1980 gggcctacgggcccgtgcac tcctcgctgt acgacctgtc ctgcatcgac acctgcgaga 2040 agaactcggtgctggaggtg atcgcctaca gcagcagcga gacccctaat cgccacgaca 2100 tgctcttggtggagccgctg aaccgactcc tgcaggacaa gtgggacaga ttcgtcaagc 2160 gcatcttctacttcaacttc ctggtctact gcctgtacat gatcatcttc accatggctg 2220 cctactacaggcccgtggat ggcttgcctc cctttaagat ggaaaaaact ggagactatt 2280 tccgagttactggagagatc ctgtctgtgt taggaggagt ctacttcttt ttccgaggga 2340 ttcagtatttcctgcagagg cggccgtcga tgaagaccct gtttgtggac agctacagtg 2400 agatgcttttctttctgcag tcactgttca tgctggccac cgtggtgctg tacttcagcc 2460 acctcaaggagtatgtggct tccatggtat tctccctggc cttgggctgg accaacatgc 2520 tctactacacccgcggtttc cagcagatgg gcatctatgc cgtcatgata gagaagatga 2580 tcctgagagacctgtgccgt ttcatgtttg tctacatcgt cttcttgttc gggttttcca 2640 cagcggtggtgacgctgatt gaagacggga agaatgactc cctgccgtct gagtccacgt 2700 cgcacaggtggcgggggcct gcctgcaggc cccccgatag ctcctacaac agcctgtact 2760 ccacctgcctggagctgttc aagttcacca tcggcatggg cgacctggag ttcactgaga 2820 actatgacttcaaggctgtc ttcatcatcc tgctgctggc ctatgtaatt ctcacctaca 2880 tcctcctgctcaacatgctc atcgccctca tgggtgagac tgtcaacaag atcgcacagg 2940 agagcaagaacatctggaag ctgcagagag ccatcaccat cctggacacg gagaagagct 3000 tccttaagtgcatgaggaag gccttccgct caggcaagct gctgcaggtg gggtacacac 3060 ctgatggcaaggacgactac cggtggtgct tcagggtgga cgaggtgaac tggaccacct 3120 ggaacaccaacgtgggcatc atcaacgaag acccgggcaa ctgtgagggc gtcaagcgca 3180 ccctgagcttctccctgcgg tcaagcagag tttcaggcag acactggaag aactttgccc 3240 tggtcccccttttaagagag gcaagtgctc gagataggca gtctgctcag cccgaggaag 3300 tttatctgcgacagttttca gggtctctga agccagagga cgctgaggtc ttcaagagtc 3360 ctgccgcttccggggagaag tgaggacgtc acgcagacag cactgtcaac actgggcctt 3420 aggagaccccgttgccacgg ggggctgctg agggaacacc agtgctctgt cagcagcctg 3480 gcctggtctgtgcctgccca gcatgttccc aaatctgtgc tggacaagct gtgggaagcg 3540 ttcttggaagcatggggagt gatgtacatc caaccgtcac tgtccccaag tgaatctcct 3600 aacagactttcaggttttta ctcactttac taaacagtkt ggatggtcag tctctactgg 3660 gacatgttaggcccttgttt tctttgattt tattcttttt tttgagacag aatttcactc 3720 ttctcacccaggctggaatg cagtggcaca attttggctc cctgcaacct ccgcctcctg 3780 gattccagcaattctcctgc ctcggcttcc caagtagctg ggattacagg cacgtgccac 3840 catgtctggctaattttttg tattttttta atagatatgg ggtttcgcca tgttggccag 3900 gctggtctcgaactcctgac ctcaggtgat ccgcccacct cggcctccca aagtgctggg 3960 attacaggtgtgagcctcca cacctggctg ttttctttga ttttattctt tttttttttt 4020 tctgtgagacagagtttcac tcttgttgcc caggctggag tgcagtggtg tgatcttggc 4080 tcactgcaacttctgcctcc cgggttcaag cgattcttct gcttcagtct cccaagtagc 4140 ttggattacaggtgagcact accacgcccg gctaattttt gtatttttaa taragacggg 4200 gtttcaccatgttggccagg ctggtctcga actcttgacc tcaggtgatc tgcccgcctt 4260 ggcctcccaaagtgctggga ttacaggtgt gagccgctgc gctcggcctt ctttgatttt 4320 atattattaggagcaaaagt aaatgaagcc caggaaaaca cctttgggaa caaactcttc 4380 ctttgatggaaaatgcagag gcccttcctc tctgtgccgt gcttgctcct cttacctgcc 4440 cgggtggtttgggggtgttg gtgtttcctc cctggagaag atgggggagg ctgtcccact 4500 cccagctctggcagaatcaa gctgttgcag cagtgccttc ttcatccttc cttacgatca 4560 atcacagtctccagaagatc agctcaattg ctgtgcaggt taaaactaca gaaccacatc 4620 ccaaaggtacctggtaagaa tgtttgaaag atcttccatt tctaggaacc ccagtcctgc 4680 ttctccgcaatggcacatgc ttccactcca tccatactgg catcctcaaa taaacagata 4740 tgtatacaataaaaaaaaaa aaaaaaaaaa rrgcggccgc tgaattctag acctgcccgg 4800 gcg 4803 4839 PRT HOMO SAPIENS UNSURE (144)(194)(198) 4 Met Lys Lys Trp Ser SerThr Asp Leu Gly Ala Ala Ala Asp Pro Leu 1 5 10 15 Gln Lys Asp Thr CysPro Asp Pro Leu Asp Gly Asp Pro Asn Ser Arg 20 25 30 Pro Pro Pro Ala LysPro Gln Leu Ser Thr Ala Lys Ser Arg Thr Arg 35 40 45 Leu Phe Gly Lys GlyAsp Ser Glu Glu Ala Phe Pro Val Asp Cys Pro 50 55 60 His Glu Glu Gly GluLeu Asp Ser Cys Pro Thr Ile Thr Val Ser Pro 65 70 75 80 Val Ile Thr IleGln Arg Pro Gly Asp Gly Pro Thr Gly Ala Arg Leu 85 90 95 Leu Ser Gln AspSer Val Ala Ala Ser Thr Glu Lys Thr Leu Arg Leu 100 105 110 Tyr Asp ArgArg Ser Ile Phe Glu Ala Val Ala Gln Asn Asn Cys Gln 115 120 125 Asp LeuGlu Ser Leu Leu Leu Phe Leu Gln Lys Ser Lys Lys His Xaa 130 135 140 ThrAsp Asn Glu Phe Lys Asp Pro Glu Thr Gly Lys Thr Cys Leu Leu 145 150 155160 Lys Ala Met Leu Asn Leu His Asp Gly Gln Asn Thr Thr Ile Pro Leu 165170 175 Leu Leu Glu Ile Ala Arg Gln Thr Asp Ser Leu Lys Glu Leu Val Asn180 185 190 Ala Xaa Tyr Thr Asp Xaa Tyr Tyr Lys Gly Gln Thr Ala Leu HisIle 195 200 205 Ala Ile Glu Arg Arg Asn Met Ala Leu Val Thr Leu Leu ValGlu Asn 210 215 220 Gly Ala Asp Val Gln Ala Ala Ala His Gly Asp Phe PheLys Lys Thr 225 230 235 240 Lys Gly Arg Pro Gly Phe Tyr Phe Gly Glu LeuPro Leu Ser Leu Ala 245 250 255 Ala Cys Thr Asn Gln Leu Gly Ile Val LysPhe Leu Leu Gln Asn Ser 260 265 270 Trp Gln Thr Ala Asp Ile Ser Ala ArgAsp Ser Val Gly Asn Thr Val 275 280 285 Leu His Ala Leu Val Glu Val AlaAsp Asn Thr Ala Asp Asn Thr Lys 290 295 300 Phe Val Thr Ser Met Tyr AsnGlu Ile Leu Ile Leu Gly Ala Lys Leu 305 310 315 320 His Pro Thr Leu LysLeu Glu Glu Leu Thr Asn Lys Lys Gly Met Thr 325 330 335 Pro Leu Ala LeuAla Ala Gly Thr Gly Lys Ile Gly Val Leu Ala Tyr 340 345 350 Ile Leu GlnArg Glu Ile Gln Glu Pro Glu Cys Arg His Leu Ser Arg 355 360 365 Lys PheThr Glu Trp Ala Tyr Gly Pro Val His Ser Ser Leu Tyr Asp 370 375 380 LeuSer Cys Ile Asp Thr Cys Glu Lys Asn Ser Val Leu Glu Val Ile 385 390 395400 Ala Tyr Ser Ser Ser Glu Thr Pro Asn Arg His Asp Met Leu Leu Val 405410 415 Glu Pro Leu Asn Arg Leu Leu Gln Asp Lys Trp Asp Arg Phe Val Lys420 425 430 Arg Ile Phe Tyr Phe Asn Phe Leu Val Tyr Cys Leu Tyr Met IleIle 435 440 445 Phe Thr Met Ala Ala Tyr Tyr Arg Pro Val Asp Gly Leu ProPro Phe 450 455 460 Lys Met Glu Lys Thr Gly Asp Tyr Phe Arg Val Thr GlyGlu Ile Leu 465 470 475 480 Ser Val Leu Gly Gly Val Tyr Phe Phe Phe ArgGly Ile Gln Tyr Phe 485 490 495 Leu Gln Arg Arg Pro Ser Met Lys Thr LeuPhe Val Asp Ser Tyr Ser 500 505 510 Glu Met Leu Phe Phe Leu Gln Ser LeuPhe Met Leu Ala Thr Val Val 515 520 525 Leu Tyr Phe Ser His Leu Lys GluTyr Val Ala Ser Met Val Phe Ser 530 535 540 Leu Ala Leu Gly Trp Thr AsnMet Leu Tyr Tyr Thr Arg Gly Phe Gln 545 550 555 560 Gln Met Gly Ile TyrAla Val Met Ile Glu Lys Met Ile Leu Arg Asp 565 570 575 Leu Cys Arg PheMet Phe Val Tyr Ile Val Phe Leu Phe Gly Phe Ser 580 585 590 Thr Ala ValVal Thr Leu Ile Glu Asp Gly Lys Asn Asp Ser Leu Pro 595 600 605 Ser GluSer Thr Ser His Arg Trp Arg Gly Pro Ala Cys Arg Pro Pro 610 615 620 AspSer Ser Tyr Asn Ser Leu Tyr Ser Thr Cys Leu Glu Leu Phe Lys 625 630 635640 Phe Thr Ile Gly Met Gly Asp Leu Glu Phe Thr Glu Asn Tyr Asp Phe 645650 655 Lys Ala Val Phe Ile Ile Leu Leu Leu Ala Tyr Val Ile Leu Thr Tyr660 665 670 Ile Leu Leu Leu Asn Met Leu Ile Ala Leu Met Gly Glu Thr ValAsn 675 680 685 Lys Ile Ala Gln Glu Ser Lys Asn Ile Trp Lys Leu Gln ArgAla Ile 690 695 700 Thr Ile Leu Asp Thr Glu Lys Ser Phe Leu Lys Cys MetArg Lys Ala 705 710 715 720 Phe Arg Ser Gly Lys Leu Leu Gln Val Gly TyrThr Pro Asp Gly Lys 725 730 735 Asp Asp Tyr Arg Trp Cys Phe Arg Val AspGlu Val Asn Trp Thr Thr 740 745 750 Trp Asn Thr Asn Val Gly Ile Ile AsnGlu Asp Pro Gly Asn Cys Glu 755 760 765 Gly Val Lys Arg Thr Leu Ser PheSer Leu Arg Ser Ser Arg Val Ser 770 775 780 Gly Arg His Trp Lys Asn PheAla Leu Val Pro Leu Leu Arg Glu Ala 785 790 795 800 Ser Ala Arg Asp ArgGln Ser Ala Gln Pro Glu Glu Val Tyr Leu Arg 805 810 815 Gln Phe Ser GlySer Leu Lys Pro Glu Asp Ala Glu Val Phe Lys Ser 820 825 830 Pro Ala AlaSer Gly Glu Lys 835 5 432 DNA HOMO SAPIENS UNSURE (83)(89) 5 gagcttctccctgcggtcaa gcagagtttc aggcagacac tggaagaact ttgccctggt 60 cccccttttaagagaggcaa gtnctcgana taggcagtct gctcagcccg aggaagttta 120 tctgcgacagttttcagggt ctctaaagcc agaggacgct gaggtcttca agagtcctgc 180 cgcttccggggagaagtgag gacgtcacgc agacagcact gtcaacactg ggccttagga 240 gaccccgttgccacgggggg ctgctgaggg aacaccagtg ctttttcagc agccttgcct 300 gggtctttgcctgcccagca tgttcccaaa tctgtgctgg acaagctgtg gggaagcgtt 360 cttgggaagcatgggggagt gatgttacat ccaaccgtca ctgtccccaa gttgaatctt 420 ccttaacaga tt432 6 65 PRT HOMO SAPIENS UNSURE (28)(30) 6 Ser Phe Ser Leu Arg Ser SerArg Val Ser Gly Arg His Trp Lys Asn 1 5 10 15 Phe Ala Leu Val Pro LeuLeu Arg Glu Ala Ser Xaa Arg Xaa Arg Gln 20 25 30 Ser Ala Gln Pro Glu GluVal Tyr Leu Arg Gln Phe Ser Gly Ser Leu 35 40 45 Lys Pro Glu Asp Ala GluVal Phe Lys Ser Pro Ala Ala Ser Gly Glu 50 55 60 Lys 65 7 3500 DNA HOMOSAPIENS 7 cagcgtcggg tgcagtttgg ccggaggttg cagtgagcag agattgccccattgcactct 60 agtctgggcg acagggtgag acacacacac acagacacac acacacacacacacacacac 120 acacaagcct aaacattcra ggccaggatg cttgacagat gttgattcataaaaatgaca 180 aaaagcacaa aatccaaaat ctcgtataag ctcagtggct gtggcagcgaggttgaagag 240 caaaggcagg ccgggcacct ggctgatgat gtgtggaccc gttgcacagcagggccccgc 300 agtgcggtgt gggtgtgggt gtgggtgggc cagtytctgc cgctcaccctattccaggga 360 cacagtctgc ttggctcttc tggactgagc catcctcatc accgagatcctccctgaatt 420 cagcccacga cagccacccc ggccgttttc cttgttctgt gtggggagggaggcagcgcg 480 gtggttatca acctcaccct gcagaggagg cacctgaggc ccagagacgaggagggatgg 540 gtctaaccca gaaccacaga tggctctgag ccgggggcct gtccaccctcccaggccgac 600 gtcagtggcc gcaggactgc ctgggccctg ctaggcctgc tcacctctgaggcctctggg 660 gtgagaggtt cagtcctgga aacacttcag ttctaggggg ctgggggcagcagcaagttg 720 gagttttggg gtaccctgct tcacagggcc cttggcaagg agggcaggtggggtctaagg 780 acaagcagtc cttactttgg gagtcaaccc cggcgtggtg gctgctgcaggttgcacact 840 gggccacaga ggatccagca aggatgaaga aatggagcag cacagacttgggggcagctg 900 cggacccact ccaaaaggac acctgcccag accccctgga tggagaccctaactccaggc 960 cacctccagc caagccccag ctctccacgg ccaagagccg cacccggctctttgggaagg 1020 gtgactcgga ggaggctttc ccggtggatt gccctcacga ggaaggtgagctggactcct 1080 gcccgaccat cacagtcagc cctgttatca ccatccagag gccaggagacggccccaccg 1140 gtgccaggct gctgtcccag gactctgtcg ccgccagcac cgagaagaccctcaggctct 1200 atgatcgcag gagtatcttt gaagccgttg ctcagaataa ctgccaggatctggagagcc 1260 tgctgctctt cctgcagaag agcaagaagc acctcacaga caacgagttcaaagaccctg 1320 agacagggaa gacctgtctg ctgaaagcca tgctcaacct gcacgacggacagaacacca 1380 ccatccccct gctcctggag atcgcgcggc aaacggacag cctgaaggagcttgtcaacg 1440 ccagctacac ggacagctac tacaagggcc agacagcact gcacatcgccatcgagagac 1500 gcaacatggc cctggtgacc ctcctggtgg agaacggagc agacgtccaggctgcggccc 1560 atggggactt ctttaagaaa accaaagggc ggcctggatt ctacttcggtgaactgcccc 1620 tgtccctggc cgcgtgcacc aaccagctgg gcatcgtgaa gttcctgctgcagaactcct 1680 ggcagacggc cgacatcagc gccagggact cggtgggcaa cacggtgctgcacgccctgg 1740 tggaggtggc cgacaacacg gccgacaaca cgaagtttgt gacgagcatgtacaatgaga 1800 ttctgatcct gggggccaaa ctgcacccga cgctgaagct ggaggagctcaccaacaaga 1860 agggaatgac gccgctggct ctggcagctg ggaccgggaa gatcggggtcttggcctata 1920 ttctccagcg ggagatccag gagcccgagt gcaggcacct gtccaggaagttcaccgagt 1980 gggcctacgg gcccgtgcac tcctcgctgt acgacctgtc ctgcatcgacacctgcgaga 2040 agaactcggt gctggaggtg atcgcctaca gcagcagcga gacccctaatcgccacgaca 2100 tgctcttggt ggagccgctg aaccgactcc tgcaggacaa gtgggacagattcgtcaagc 2160 gcatcttcta cttcaacttc ctggtctact gcctgtacat gatcatcttcaccatggctg 2220 cctactacag gcccgtggat ggcttgcctc cctttaagat ggaaaaaactggagactatt 2280 tccgagttac tggagagatc ctgtctgtgt taggaggagt ctacttctttttccgaggga 2340 ttcagtattt cctgcagagg cggccgtcga tgaagaccct gtttgtggacagctacagtg 2400 agatgctttt ctttctgcag tcactgttca tgctggccac cgtggtgctgtacttcagcc 2460 acctcaagga gtatgtggct tccatggtat tctccctggc cttgggctggaccaacatgc 2520 tctactacac ccgcggtttc cagcagatgg gcatctatgc cgtcatgatagagaagatga 2580 tcctgagaga cctgtgccgt ttcatgtttg tctacgtcgt cttcttgttcgggttttcca 2640 cagcggtggt gacgctgatt gaagacggga agaatgactc cctgccgtctgagtccacgt 2700 cgcacaggtg gcgggggcct gcctgcaggc cccccgatag ctcctacaacagcctgtact 2760 ccacctgcct ggagctgttc aagttcacca tcggcatggg cgacctggagttcactgaga 2820 actatgactt caaggctgtc ttcatcatcc tgctgctggc ctatgtaattctcacctaca 2880 tcctcctgct caacatgctc atcgccctca tgggtgagac tgtcaacaagatcgcacagg 2940 agagcaagaa catctggaag ctgcagagag ccatcaccat cctggacacggagaagagct 3000 tccttaagtg catgaggaag gccttccgct caggcaagct gctgcaggtggggtacacac 3060 ctgatggcaa ggacgactac cggtggtgct tcagggtgga cgaggtgaactggaccacct 3120 ggaacaccaa cgtgggcatc atcaacgaag acccgggcaa ctgtgagggcgtcaagcgca 3180 ccctgagctt ctccctgcgg tcaagcagag tttcaggcag acactggaagaactttgccc 3240 tggtccccct tttaagagag gcaagtgctc gagataggca gtctgctcagcccgaggaag 3300 tttatctgcg acagttttca gggtctctga agccagagga cgctgaggtcttcaagagtc 3360 ctgccgcttc cggggagaag tgaggacgtc acgcagacag cactgtcaacactgggcctt 3420 aggagacccc gttgccacgg ggggctgctg agggaacacc agtgctctgtcagcagcctg 3480 gcctggtctg tgcctgccca 3500 8 839 PRT HOMO SAPIENS 8 MetLys Lys Trp Ser Ser Thr Asp Leu Gly Ala Ala Ala Asp Pro Leu 1 5 10 15Gln Lys Asp Thr Cys Pro Asp Pro Leu Asp Gly Asp Pro Asn Ser Arg 20 25 30Pro Pro Pro Ala Lys Pro Gln Leu Ser Thr Ala Lys Ser Arg Thr Arg 35 40 45Leu Phe Gly Lys Gly Asp Ser Glu Glu Ala Phe Pro Val Asp Cys Pro 50 55 60His Glu Glu Gly Glu Leu Asp Ser Cys Pro Thr Ile Thr Val Ser Pro 65 70 7580 Val Ile Thr Ile Gln Arg Pro Gly Asp Gly Pro Thr Gly Ala Arg Leu 85 9095 Leu Ser Gln Asp Ser Val Ala Ala Ser Thr Glu Lys Thr Leu Arg Leu 100105 110 Tyr Asp Arg Arg Ser Ile Phe Glu Ala Val Ala Gln Asn Asn Cys Gln115 120 125 Asp Leu Glu Ser Leu Leu Leu Phe Leu Gln Lys Ser Lys Lys HisLeu 130 135 140 Thr Asp Asn Glu Phe Lys Asp Pro Glu Thr Gly Lys Thr CysLeu Leu 145 150 155 160 Lys Ala Met Leu Asn Leu His Asp Gly Gln Asn ThrThr Ile Pro Leu 165 170 175 Leu Leu Glu Ile Ala Arg Gln Thr Asp Ser LeuLys Glu Leu Val Asn 180 185 190 Ala Ser Tyr Thr Asp Ser Tyr Tyr Lys GlyGln Thr Ala Leu His Ile 195 200 205 Ala Ile Glu Arg Arg Asn Met Ala LeuVal Thr Leu Leu Val Glu Asn 210 215 220 Gly Ala Asp Val Gln Ala Ala AlaHis Gly Asp Phe Phe Lys Lys Thr 225 230 235 240 Lys Gly Arg Pro Gly PheTyr Phe Gly Glu Leu Pro Leu Ser Leu Ala 245 250 255 Ala Cys Thr Asn GlnLeu Gly Ile Val Lys Phe Leu Leu Gln Asn Ser 260 265 270 Trp Gln Thr AlaAsp Ile Ser Ala Arg Asp Ser Val Gly Asn Thr Val 275 280 285 Leu His AlaLeu Val Glu Val Ala Asp Asn Thr Ala Asp Asn Thr Lys 290 295 300 Phe ValThr Ser Met Tyr Asn Glu Ile Leu Ile Leu Gly Ala Lys Leu 305 310 315 320His Pro Thr Leu Lys Leu Glu Glu Leu Thr Asn Lys Lys Gly Met Thr 325 330335 Pro Leu Ala Leu Ala Ala Gly Thr Gly Lys Ile Gly Val Leu Ala Tyr 340345 350 Ile Leu Gln Arg Glu Ile Gln Glu Pro Glu Cys Arg His Leu Ser Arg355 360 365 Lys Phe Thr Glu Trp Ala Tyr Gly Pro Val His Ser Ser Leu TyrAsp 370 375 380 Leu Ser Cys Ile Asp Thr Cys Glu Lys Asn Ser Val Leu GluVal Ile 385 390 395 400 Ala Tyr Ser Ser Ser Glu Thr Pro Asn Arg His AspMet Leu Leu Val 405 410 415 Glu Pro Leu Asn Arg Leu Leu Gln Asp Lys TrpAsp Arg Phe Val Lys 420 425 430 Arg Ile Phe Tyr Phe Asn Phe Leu Val TyrCys Leu Tyr Met Ile Ile 435 440 445 Phe Thr Met Ala Ala Tyr Tyr Arg ProVal Asp Gly Leu Pro Pro Phe 450 455 460 Lys Met Glu Lys Thr Gly Asp TyrPhe Arg Val Thr Gly Glu Ile Leu 465 470 475 480 Ser Val Leu Gly Gly ValTyr Phe Phe Phe Arg Gly Ile Gln Tyr Phe 485 490 495 Leu Gln Arg Arg ProSer Met Lys Thr Leu Phe Val Asp Ser Tyr Ser 500 505 510 Glu Met Leu PhePhe Leu Gln Ser Leu Phe Met Leu Ala Thr Val Val 515 520 525 Leu Tyr PheSer His Leu Lys Glu Tyr Val Ala Ser Met Val Phe Ser 530 535 540 Leu AlaLeu Gly Trp Thr Asn Met Leu Tyr Tyr Thr Arg Gly Phe Gln 545 550 555 560Gln Met Gly Ile Tyr Ala Val Met Ile Glu Lys Met Ile Leu Arg Asp 565 570575 Leu Cys Arg Phe Met Phe Val Tyr Val Val Phe Leu Phe Gly Phe Ser 580585 590 Thr Ala Val Val Thr Leu Ile Glu Asp Gly Lys Asn Asp Ser Leu Pro595 600 605 Ser Glu Ser Thr Ser His Arg Trp Arg Gly Pro Ala Cys Arg ProPro 610 615 620 Asp Ser Ser Tyr Asn Ser Leu Tyr Ser Thr Cys Leu Glu LeuPhe Lys 625 630 635 640 Phe Thr Ile Gly Met Gly Asp Leu Glu Phe Thr GluAsn Tyr Asp Phe 645 650 655 Lys Ala Val Phe Ile Ile Leu Leu Leu Ala TyrVal Ile Leu Thr Tyr 660 665 670 Ile Leu Leu Leu Asn Met Leu Ile Ala LeuMet Gly Glu Thr Val Asn 675 680 685 Lys Ile Ala Gln Glu Ser Lys Asn IleTrp Lys Leu Gln Arg Ala Ile 690 695 700 Thr Ile Leu Asp Thr Glu Lys SerPhe Leu Lys Cys Met Arg Lys Ala 705 710 715 720 Phe Arg Ser Gly Lys LeuLeu Gln Val Gly Tyr Thr Pro Asp Gly Lys 725 730 735 Asp Asp Tyr Arg TrpCys Phe Arg Val Asp Glu Val Asn Trp Thr Thr 740 745 750 Trp Asn Thr AsnVal Gly Ile Ile Asn Glu Asp Pro Gly Asn Cys Glu 755 760 765 Gly Val LysArg Thr Leu Ser Phe Ser Leu Arg Ser Ser Arg Val Ser 770 775 780 Gly ArgHis Trp Lys Asn Phe Ala Leu Val Pro Leu Leu Arg Glu Ala 785 790 795 800Ser Ala Arg Asp Arg Gln Ser Ala Gln Pro Glu Glu Val Tyr Leu Arg 805 810815 Gln Phe Ser Gly Ser Leu Lys Pro Glu Asp Ala Glu Val Phe Lys Ser 820825 830 Pro Ala Ala Ser Gly Glu Lys 835

What is claimed is:
 1. An isolated polynucleotide comprising thenucleotide sequence of SEQ ID NO:1.
 2. An isolated polynucleotideconsisting of the nucleotide sequence of SEQ ID NO:1.
 3. An isolatedpolynucleotide that is RNA corresponding to the isolated polynucleotideset forth in SEQ ID NO:1.
 4. An isolated polynucleotide that is fullycomplementary to the nucleotide sequence of SEQ ID NO:1.
 5. An isolatedpolynucleotide comprising the nucleotide sequence of SEQ ID NO:7.
 6. Anisolated polynucleotide comprising the nucleotide sequence of SEQ IDNO:3.